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首页> 外文期刊>Plant Biotechnology Reports >Overexpression of poplar GSTU51 confers selective tolerance to both mercury and methyl viologen but not to CDNB or cadmium in transgenic poplars
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Overexpression of poplar GSTU51 confers selective tolerance to both mercury and methyl viologen but not to CDNB or cadmium in transgenic poplars

机译:杨树GSTU51的过表达赋予转基因杨树对汞和甲基紫精的选择性耐受性,但对CDNB或镉不具有选择性耐受性

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摘要

Glutathione S-transferases belong to a large ancient gene family and are thought to be one of the effective detoxification systems. To elucidate the function of the gene in poplar, a tau class gst gene (PatgGSTU51) was cloned from poplar cell suspension cDNA library and its expression was examined. The gene was not expressed in normal conditions, but significantly induced by toxic heavy metals like cadmium and inorganic mercury. However, the highest expression was observed when treated with its synthetic substrate, 1-chloro-2,4-dinitrobenzene (CDNB). Several transgenic poplar lines harboring a chimeric p35S-PatgGSTU51 were developed to understand its function in plant defense. Real-time quantitative PCR, western blot and cellular GST activities consistently showed the transgene was highly expressed in the transgenic lines. The transgenic lines showed increased tolerance to methyl viologen as well as to inorganic mercury but not to cadmium. Furthermore, they did not show any significant tolerance against CDNB, the electrophilic substrate of GST isozymes. Thus, the results suggest that, although PatgGSTU51 is induced by a number of stress agents, it confers a selective tolerance to the toxicity of heavy metal mercury over those of other stress agents and also to oxidation stress caused by MV. Tolerance to CDNB that induces the gene to high level may require such an extra supplemental gene action as vacuolar sequestration.
机译:谷胱甘肽S-转移酶属于一个古老的大基因家族,被认为是有效的排毒系统之一。为了阐明该基因在杨树中的功能,从杨树细胞悬液cDNA文库中克隆了tau类gst基因(PatgGSTU51),并检查了其表达。该基因在正常条件下不表达,但由镉和无机汞等有毒重金属显着诱导。但是,当用其合成底物1-氯-2,4-二硝基苯(CDNB)处理时,观察到最高的表达。为了了解其在植物防御中的功能,开发了几种带有嵌合p35S-PatgGSTU51的转基因杨树品系。实时定量PCR,Western印迹和细胞GST活性一致表明该转基因在转基因品系中高度表达。转基因品系显示出对甲基紫精和无机汞的耐受性增强,但对镉的耐受性却增强。此外,它们对CDNB(GST同工酶的亲电底物)没有表现出明显的耐受性。因此,结果表明,尽管PatgGSTU51是由多种胁迫剂诱导的,但它赋予了重金属汞毒性超过其他胁迫剂的选择性耐受性以及对MV引起的氧化胁迫的选择性耐受性。对诱导基因高水平表达的CDNB的耐受性可能需要液泡螯合等额外的补充基因作用。

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